Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields
In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS<sup>®</sup>D4C depths 10 and 20 µm, ISAS<sup>®</sup>D4C Leja 20 and dro...
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doaj-cb97c2638e994d2eaf63f89e116d45e02020-11-25T04:06:12ZengMDPI AGAnimals2076-26152020-10-01101993199310.3390/ani10111993Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and FieldsSabrina Gacem0Jaime Catalán1Anthony Valverde2Carles Soler3Jordi Miró4Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, 08193 Bellaterra, SpainEquine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, 08193 Bellaterra, SpainCosta Rica Institute of Technology, School of Agronomy, San Carlos Campus, 223-21001 Alajuela, Costa RicaDepartamento de Biología Celular, Biología Funcional y Antropología Física, Universitat de València, 46100 Burjassot, Valencia, SpainEquine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, 08193 Bellaterra, SpainIn order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS<sup>®</sup>D4C depths 10 and 20 µm, ISAS<sup>®</sup>D4C Leja 20 and drop displacement chamber (DDC), Spermtrack<sup>®</sup> (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 10<sup>6</sup> sperm/mL offers the best choice for donkey computerised sperm motility analysis.https://www.mdpi.com/2076-2615/10/11/1993frame ratedrop displacement chamberscapillary loaded chamberschamber depthfieldsperm dilution |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sabrina Gacem Jaime Catalán Anthony Valverde Carles Soler Jordi Miró |
spellingShingle |
Sabrina Gacem Jaime Catalán Anthony Valverde Carles Soler Jordi Miró Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields Animals frame rate drop displacement chambers capillary loaded chambers chamber depth field sperm dilution |
author_facet |
Sabrina Gacem Jaime Catalán Anthony Valverde Carles Soler Jordi Miró |
author_sort |
Sabrina Gacem |
title |
Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields |
title_short |
Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields |
title_full |
Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields |
title_fullStr |
Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields |
title_full_unstemmed |
Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields |
title_sort |
optimization of casa-mot analysis of donkey sperm: optimum frame rate and values of kinematic variables for different counting chamber and fields |
publisher |
MDPI AG |
series |
Animals |
issn |
2076-2615 |
publishDate |
2020-10-01 |
description |
In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS<sup>®</sup>D4C depths 10 and 20 µm, ISAS<sup>®</sup>D4C Leja 20 and drop displacement chamber (DDC), Spermtrack<sup>®</sup> (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 10<sup>6</sup> sperm/mL offers the best choice for donkey computerised sperm motility analysis. |
topic |
frame rate drop displacement chambers capillary loaded chambers chamber depth field sperm dilution |
url |
https://www.mdpi.com/2076-2615/10/11/1993 |
work_keys_str_mv |
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